agilent solutions for preimplantation genetic testing · 5 rechitsky s., fertil steril. 2015...
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Agilent solutions for preimplantation genetic testingMake informed choices with technologies that fit your needs
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Agilent is your trusted partner for embryo testing and ranking. Our mission is to accelerate informed decisions by providing tools and solutions that address laboratory needs in terms of throughput, effectiveness, completeness and ease of use.
Agilent is innovating the path for embryo testing and ranking
The first applications of in vitro fertilization (IVF) date back 40 years1. Since then, the introduction of new procedures and technologies have drastically changed the way we approach IVF. In the early 1990s, the first successful clinical preimplantation genetic testing (PGT) was applied to eliminate the transmission of X-linked conditions and cystic fibrosis transmembrane conductance regulator (CFTR) mutations2,3. Today, PGT is commonly used to test a few cells from embryos before embryo transfer to detect aneuploidy (PGT-A), translocations (PGT-SR), and/or more than 400 monogenic disorders (PGT-M). It has been estimated that approximately 100,000 PGT cycles have been performed worldwide over the past 23 years, the vast majority being PGT-A (www.pgdis.org).
Brief history of IVF
The benefits of embryo testingTo increase the success of IVF procedures, embryo testing4,5 and ranking is a routine clinical procedure in cytogenetic laboratories and IVF clinics prior to implantation. The benefits include:
– Reduction in miscarriage rate
– Increased pregnancy rate
– Increased likelihood of healthy birth
– Reduced risk of multiple pregnancies
– Optimization of time and resources
Informed testing, that can be detected with available technologies Current PGT nomenclature highlights the different applications of preimplantation genetic testing of embryos. The new definitions highlight all the genetic anomalies tested:
By providing highly accurate and comprehensive genetic data, Agilent enables fertility specialists to make better-informed decisions on embryo ranking and selection—thereby increasing the chances of a successful and healthy pregnancy. In addition, Agilent’s product portfolio provides a full spectrum of solutions linked to human reproductive genetics.
Agilent provides two complementary integrated solutions for the detection of aneuploidies, chromosomal rearrangements and single gene disorders or a combination thereof.
– GenetiSure Pre-Screen kits
– CGH array platform solution
– OnePGT Solution - Genome-wide next-generation sequencing (NGS) solution
Prenatal and postnatal genome analysis with a full suite of technologies from Agilent
NIPTClarigo* is a unique, NGS-based, CE-marked noninvasive prenatal test (NIPT) solution validated for detection of trisomy 13, 18, 21 with optional gender calling in cell-free DNA from maternal blood.
CGHIndustry-leading catalog and custom CGH microarrays are able to detect whole genome CNVs and absence of heterozygosity (AOH) in a single experiment using blood, CVS, or amniotic fluid as starting samples. The CGH and CGH+SNP platform offers real comparative genomic data due to the 2-color approach, and unlimited flexibility in terms of format and content.
NGSAgilent’s market-leading hybrid capture method, SureSelect, offers NGS target enrichment solutions including custom & catalog panels to analyze a few genes to full exomes. SureMASTR catalog panels are based on multiplex amplification of selected genomic targets through a simple PCR assay. Panels are focused on specific diseases or conditions (e.g. CFTR, BRCA) and come with MASTR Reporter, a dedicated data analysis tool.
VS.
OR
PGT-SR PGT-A
PGT-M
G T T A AG
GC CA A
G T T A AG
GC CA G
G T T A AG
GC CA T
SNP
PGT-A
PGT-SR
PGT-M
GenetiSure
Pre-Screen
OnePG
T
* The product referenced above is not available for sale in all countries or jurisdictions. Please contact your local sales representative for additional information.
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Agilent OnePGT solution is a universal genome-wide NGS-based solution that integrates preimplantation genetic testing (PGT) for single gene disorders (PGT-M), translocations (PGT-SR) and aneuploidies (PGT-A) in a single workflow with verified automated calling.OnePGT is developed to help fertility specialists and laboratories achieve comprehensive insights to guide embryo ranking. This all-in-one solution enables labs to concurrently test for three PGT applications using a single biopsy sample.
OnePGT: Comprehensive insight for every IVF transfer
Sample collection(Day 3 to day 5)
NGSlibrary prep
NGSAutomated
data analysisDNA
DNA from parents & phasing references
One sample, one workflow, comprehensive insights
Multiple Displacement Amplification
(MDA)
For Research Use Only. Not for use in diagnostic procedures.
Single gene disorders Translocations Aneuploidy testing
VS.
VS. VS.
PGT-M PGT-SR PGT-A
VS.
OR
REPLI-g Single Cell kit
Agilent Alissa OnePGT software
Agilent OnePGT Library Prep kit
Agilent OnePGT solution
Sample collection(Day 3 to day 5)
NGSlibrary prep
NGSAutomated
data analysisDNA
DNA from parents & phasing references
One sample, one workflow, comprehensive insights
Multiple Displacement Amplification
(MDA)
For Research Use Only. Not for use in diagnostic procedures.
Single gene disorders Translocations Aneuploidy testing
VS.
VS. VS.
PGT-M PGT-SR PGT-A
VS.
OR
REPLI-g Single Cell kit
Agilent Alissa OnePGT software
Agilent OnePGT Library Prep kit
Agilent OnePGT solution
Sample collection(Day 3 to day 5)
NGSlibrary prep
NGSAutomated
data analysisDNA
DNA from parents & phasing references
One sample, one workflow, comprehensive insights
Multiple Displacement Amplification
(MDA)
For Research Use Only. Not for use in diagnostic procedures.
Single gene disorders Translocations Aneuploidy testing
VS.
VS. VS.
PGT-M PGT-SR PGT-A
VS.
OR
REPLI-g Single Cell kit
Agilent Alissa OnePGT software
Agilent OnePGT Library Prep kit
Agilent OnePGT solution
OnePGT
1 Genome-wide NGS
Maximize efficiency in the selection of embryo’s
Contribute to successful pregnancies
Have peace of mind in running your assay
2 Comprehensive automatic calling
3 Single, universal workflow
4 Verified on clinical samples
5 Embedded QC metrics
6 On-boarding program
thanks to … enables you to …
Advantages
Comprehensive insights
— Facilitates multiple requests on same biopsy
— Complete report to improve cycle management
Easy
— Eliminates test development
— One workflow for PGT-M/SR/A
— Automated calling, including segmentals
Reliable
— Verified on clinical samples
— Embedded QC metrics
— Scalable and flexible
Fast implementation
— Optimized workflow
— Proven performance
— Onboarding program
Figure 2. OnePGT can be used to determine the genetic profile of an embryo with regard to an inherited single gene disorder (PGT-M) or translocation (PGT-SR) as well as chromosomal copy number aberrations (PGT-A) using a single optimized NGS workflow.
GenetiSure Pre-Screen Complete kit with QIAGEN REPLI-g is a cost-effective and a highly sensitive array CGH solution. The assay includes all the reagents and the software with dedicated analysis workflow.Microarrays can be applied in PGT when high resolution and fast results are requested. Variable array formats enable scalable throughput using blastomere or trophectoderm biopsies from fresh or frozen embryos.
GenetiSure Pre-Screen solution – rapid and reliable identification of aneuploidies and chromosomal rearrangements
Protocol workflow
20 minSample preparation
20 minPurification
80 minLabeling
2 hrHybridization
140 minREPLI-g WGA
35 minPreparation for hybridization
45 min
7 hr 40 minTotal:
Washing, scanning, analysis
Figure 1. A) Thanks to higher data quality linked to MDA amplification, the new kit enables calling with higher confidence small aberrations, in the range of 1-5 Mb, above an example of two deletion < 2 Mb detected with the platform. B) Working with single cells provides extra flexibility to the solution, allowing to work with day 3 embryo biopsy if needed. The pre-screen complete platform is robust enough to call with confidence aberrations > 5 Mb also starting from such limited starting material. Above is an example of a gain and a loss detected starting from a single cell (cell line GM14485). C) The ability to call mosaic is extremely important to properly rank embryo quality. We mixed two cell lines both carrying a gain of approx. 30 Mb to test ability of the assay to detect low level mosaic down to 20%.
Advantages
— Easy Based on Agilent CGH platform
— High resolution Ability to call aberrations down to >1 Mb
— Fast From sample to results in less than 8 hours
— Low-cost Cost-effective, starting from 6 samples/run
A. B. C.
4
6 7
Sample collection(Day 3 to day 5)
Highlights of MDA
REPLI-g Single Cell kit
Benefits of MDA
CN information is retained
Strong strand displacement activity & high fidelity DNA synthesis
Very low DNA fragmentation, longer fragments
Secondary structures resolved for easier amplification
Minimum bias in genome coverage
Non PCR based amplification
Optimized Φ29 enzyme
Optimized alkaline incubation
Single cell buffer system
Average product length 10 Kb
– Results in 8 h – Resolution down to less than 2 Mb– Cost-effective (start with as few as 6 samples/run)
– Flexible genome-wide NGS-based solution with onboarding program– Universal workflow for all PGT applications– Comprehensive automated calling verified on clinical samples
Automateddata analysis
NGSlibrary prep
NGSData analysis HybridizationWash & scan Labeling
CGH NGS
Comprehensive insights to facilitate embryo ranking and detection of monogenic disorders
Two technologies – Comprehensive preimplantation genetic testing
Agilent is the only commercial manufacturer offering two reliable technologies (CGH and NGS) for universal genome-wide comprehensive preimplantation genetic testing. We offer a complete solution with reagents and software for both CGH and NGS workflows, including sample preparation, quality control and data analysis.
Both technologies include the same DNA amplification protocol, based on the QIAGEN REPLI-g kit MDA technology. This protocol gives you flexibility to collect your embryo sample, perform Multiple Displacement Amplification (MDA), and then choose between the CGH or NGS workflow based on your needs.
For Research Use Only. Not for use in diagnostic procedures.
This information is subject to change without notice.
PR7000-2219 © Agilent Technologies, Inc. 2019 Published in the USA, April 2, 2019 5994-0875EN
1 Steptoe PC & Edwards RG 1978 Birth after the reimplantation of a human embryo. Lancet 2 366. (https://doi.org/10.1016/S0140-6736(78)92957- 4)
2 A review of the modern approach to in vitro fertilization Susan M. Maxwell, MD *, James A. Grifo, MD, PhD
3 Chromosomal analysis in IVF: just how useful is it? Darren K Griffin1 and Cagri Ogur2,3
1School of Biosciences, Centre for Interdisciplinary Studies of Reproduction, University of Kent, Canterbury, UK, 2Bahceci Genetic Diagnosis Center, İstanbul, Turkey and
3Department of Bioengineering, Yildiz Technical University, İstanbul, Turkey4 Goldman KN., J Genet Couns. 2016 Dec;25(6):1327-1337;5 Rechitsky S., Fertil Steril. 2015 Feb;103(2):503-12
Product Description Part number
Agilent OnePGT solution Includes reagents for genome-wide amplification and library preparation, software and cloud services
G9426AA
Agilent OnePGT solution without REPLI-g Includes reagents for library preparation, software and cloud services G9427AA
Agilent OnePGT HOT workshop Hands-On-Training workshop in Agilent facility to train customers on OnePGT workflow and data analysis
R2600A
OnePGT ordering information
GenetiSure Pre-Screen kit ordering information
† The SureTag Complete DNA Labeling kit includes 50 SureTag purification columns. Use p/n 5190-3391 to purchase additional columns needed to process 8x microarray.
* G5761AA SureScan Dx available in select countries.
Product number Product Description
G9500A GenetiSure Pre-Screen Complete kit 8x60K Array and reagents to analyze 42 samples and 6 controls
G9501A GenetiSure Pre-Screen Complete kit 4x180K Array and reagents to analyze 32 samples and 12 controls
G9502A GenetiSure Pre-Screen Amplification & Labeling kit Amplification and labeling reagents to process 48 samples (samples and controls)
G5963A GenetiSure Pre-Screen Array kit 8x60K 3 slides – 48 samples
G5962A GenetiSure Pre-Screen Array kit 4x180K 3 slides – 24 samples
G5972A GenetiSure Pre-Screen kit 8x60K - X/Y 3 slides – 48 samples
G5973A GenetiSure Pre-Screen kit 8x60K - X/Y 3 slides – 24 samples
Product number Product
5190-4240SureTag Complete DNA Labeling kit† (including Agilent
M/F Reference, 100 rxn 8x, 50 rxn 4x)
5190-3393 Human Cot-1 DNA
Low 5188-5220
High 5188-5380
Agilent Oligo aCGH Hybridization kit to process 25 (low)
or 100 (high) slides
5188-5226 Agilent Oligo aCGH Wash Buffer 1 and 2 Set
G2534-60018 Hybridization Chamber Gasket Slide kit 8-pack, 3 gasket slides
G2534-60011 Hybridization Chamber Gasket Slide kit 4-pack, 5 gasket slides
Product number Product
G2534A Hybridization chamber, stainless
G2545A Hybridization oven
G2530-60029 Hybridization oven rotator rack
G4900DA SureScan microarray scanner*
Agilent CytoGenomics software v5.0 license (free)
Download: https://www.agilent.com/en/download-agilent-cytogenomics-software
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